Native isolates T. citrinoviride (T33) and Trichoderma sp.anamorph ofHypocrea semiorbis (T15)wereisolated from rhizosphere soil of corn in Yemen and selected for their high biocontrol activity against different soil-borne plant pathogens.
F. oxysporum (AUMC 208) was collected from Mycological Center, Faculty of Science, Assiut University, Assiut, Egypt.
Soil used through this experiment was clay soil, sand and peat moss at a ratio of 3:1:1 (w/w) then autoclaved for three successive days at 121°C for one hour/day. Pots of 15 cm ×10 cm diameter were surface sterilized with 1% sodium hypochlorite and were filled with autoclaved soil [11, 12]. Seeds of tomato were surface sterilized with 1% sodium hypochlorite for 2 min, rinsed three times in sterile distilled water and air dried .
Fusarium oxysporum inocula was prepared for incula by growing on PDA and incubated for 7 days at 28°C. The suspension for inoculation was prepared by pouring 30 ml of sterile distilled water into each of petri dishes containing F. oxysporum culture and the conidia were dislodged with a cell spreader, filtered through cheesecloth and counted with a haemacytometer. It was set to 1x106 conidia/ml .
Pathogenicity test of F. oxysporum was carried out on tomato variety (Solanum lycopersicum L., cvs. ‘Rio grande, Faten) according to Jasnic ., et al. . Pathogenicity test was tested by sowing of tomato seeds in artificially infected soil by F. oxysporum. Control plants were sown in soil with sterile distilled water. The severity of the Fusarium wilt was assessed from 2 weeks of inoculation up to 45 days. Disease incidence was determined as reported by Khanna., et al. :
n= number of plants showing wilts symptoms
N= Total number of plant sampled
The selected antagonists (T15 and T33) were tested for their ability to control the Fusarium wilt of tomato as well as their effect on tomato plant growth under greenhouse conditions. Each pot was taken with sterilized soil. Soil was infested with conidial suspension (30 ml) of F. oxysporum (1x106 conidia/ml). Simultaneously after 2 days of pathogen inoculation, soils were inoculated with (30 ml) of Trichoderma isolates (1x107 conidia/ml) and then pots were watered for 7 days before sowing. Ten tomato seeds were sown in each pot, three pot replicate for each treatment arranged in completed randomized design. Pots were kept under greenhouse condition till the end of the experiment . The details of total 5 treatments as follow:
T (1): Control (without Fusarium and Trichoderma)
T (2): F. oxysporum
T (3): Trichoderma sp.anamorph of H. semiorbis+ F. oxysporum
T (4): T. citrinoviride + F. oxysporum
T (5): Trichoderma sp.anamorph of H. semiorbis +T. citrinoviride+ F. oxysporum
- no symptoms
- chlorosis and wilting of the first branches
- chlorosis and wilting of second and third branches
- chlorosis above third, second and third branches may be lost
- chlorosis and partial desiccation
- complete death of plant
Statistical analysis was performed in order to determine the effect of treatments on observed parameters. Significance of treatments was tested by One-way Analysis of Variance (ANOVA) and Tukey’s test (at P< 0.05) was applied for the differences in mean values. All the statistical analyses were completed using Graphpad 6.01 Statistics.
|Treatments||Plant length (cm)||Fresh weight (g)||Dry weight (g)||No. of leaves|
|T3||F + T15||5.60||15.76||2.85||18.66||0.38||2.62||20.68|
|T4||F + T33||5.75||16.64||2.90||21.12||0.44||3.01||22.62|
We thank Faculty of Science, University of Sana’a for their support during this investigation.
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